CN112146362B

CN112146362B - Apparatus for maintaining a controlled environment

Info

Publication number: CN112146362B
Application number: CN202010915934.3A
Authority: CN
Inventors: B.J.奈特
Original assignee: Gen Probe Inc
Current assignee: Gen Probe Inc
Priority date: 2015-08-03
Filing date: 2016-08-02
Publication date: 2022-06-03
Anticipated expiration: 2036-08-02
Also published as: US11668525B2; KR20200096698A; US20210310734A1; US11035613B2; US20190390903A1; CN107923706A; EP3949732A1; EP3332200B1; US20180209731A1; US10443935B2; KR102408787B1; USD940305S1; KR20180035890A; KR102296720B1; CN107923706B; EP3332200A1; CN112146362A; WO2017023934A1

Abstract

The present application describes lyophilization kits and methods of use thereof. In various embodiments, the lyophilization nest is configured to support one or more reservoirs that each hold one or more substances within the interior space of the lyophilization nest. The interior space may be in fluid communication with the exterior of the lyophilization nest via one or more vent holes extending through a surface of the lyophilization nest. The one or more vents each have a corresponding sealing element configured to selectively form an airtight seal within the vent such that a controlled environment may be maintained within the interior space when the environmental condition surrounding the lyophilization nest is not a lyophilization condition. One or more sealing elements may be operated to form an air-tight seal by pressing the sealing elements into corresponding vent holes when the lyophilization nest is in the sealed lyophilizer.

Description

Apparatus for maintaining a controlled environment

This application is a divisional patent application of the invention patent application having an application date of 2016, 8/2, application number of 201680046000.2, entitled "apparatus for maintaining controlled environment".

Cross Reference to Related Applications

This application claims priority to U.S. provisional application No. 62/200,370 filed on 3/8/2015, which is incorporated by reference into this application.

Background

Lyophilization is a process used to remove water from substances so that the substances can be stored for longer periods of time without deterioration of the material. Typically, these substances are of biological or synthetic origin, which may include antibiotics, drugs, chemicals, serum, vaccines, cells, tissues, proteins, and/or nucleic acids.

By placing a single vial or multi-well plate containing one or more substances into a lyophilizer, the substances can be lyophilized, with the lyophilizer creating and maintaining a controlled environment therein during the lyophilization process. Environmental factors for freeze dryer control include, but are not limited to, temperature, air pressure/vacuum, humidity, and/or gas content. After lyophilization is complete, the seal on the lyophilizer is removed, thus allowing the controlled environment within the lyophilizer to be diluted by the air surrounding the lyophilizer, which can be detrimental to the lyophilized material. The main result of contacting the lyophilized substance with air from outside the lyophilizer is that the air usually contains moisture (e.g. water vapor) that can rehydrate the lyophilized substance (at least partially). Depending on factors such as time of year, air conditioning of the room, number of people in the room, and exposure time, the lyophilized material that comes into contact with air from outside the lyophilizer can undergo 40% to 70% rehydration. The introduction of moisture into the lyophilized material can adversely affect the stability of the material. If the substance is not to be used immediately, the wells of the vials and/or multi-well plates can be sealed by placing a foil or other sealing material over the wells and/or vials, thereby sealing and isolating the lyophilized substance from the air. After the plate has been removed from the lyophilizer, the seal provides a barrier between the ambient conditions surrounding the plate and the substance located within the wells of each vial or plate. Thus, the seal prevents rehydration of the substance by absorption of water vapor from the atmosphere. The sealed vials and/or plates containing the one or more substances may then be stored or sold commercially for later use. The seal may then be broken so that the stored contents of the vial and/or one or more of the individual wells may be accessed during use.

The process of sealing the reservoir after removal from the lyophilizer exposes the individual substances to adverse environmental conditions for the following periods of time: the time period between removal of the reservoir from the lyophilizer until the reservoir is sealed. While this exposure time may vary depending on the number of vials and/or wells of a multi-well plate to be sealed, the number of technicians available to seal the vials and/or wells, the transport distance between the lyophilizer and the sealing location, and other factors experienced during the lyophilization process, the exposure time of the lyophilized material in a high throughput facility may be between 4-5 hours. Thus, the lyophilized material in the wells of each vial or multiwell plate may be exposed to uncontrolled and potentially adverse environmental conditions, including moisture, prior to sealing.

The time during which the lyophilized material is exposed to these uncontrolled environmental conditions for an extended period of time during the time period between removal of the reservoir from the lyophilizer until the reservoir is sealed can be reduced as follows: immediately after removal from the lyophilizer, the reservoirs were placed into a glove box (e.g., Purelab HE 4GB 2500Glovebox, available from Innovating Technology, inc. However, when using a glove box, the operation of the reservoir may be cumbersome. Furthermore, during the transfer of the reservoir from the lyophilizer to the glove box, the wet vapor exposure time may still be non-negligible and may lead to hygroscopic freeze-dried material. Furthermore, the environment inside the glove box cannot be made the same as inside the freeze dryer itself, so the freeze-dried substance is still exposed to adverse humidity levels for a long time before being sealed.

The freeze dryer can be operated in a drying chamber with low humidity. Upon removal of the reservoir from the lyophilizer, the material can only be exposed to the drying chamber before sealing. However, drying chambers require a precise climate control system to maintain a low humidity in the interior space sized to accommodate at least one operator. However, the humidity level of the drying chamber is also different from the humidity level in the freeze dryer, and the presence of the operator alone in the drying chamber further adversely affects the environmental conditions. Drying rooms are expensive to maintain, prone to contamination, and cumbersome to operate because the conditions can be dangerous for individuals working in these rooms, and thus the amount of time an individual can work in these rooms can be limited.

A large single vial with a stopper storing a single substance may be configured for stopper engagement when the single vial is located within a sealed lyophilizer. However, such single vials are difficult to manage, bulky, difficult to store, and generally not compatible with laboratory equipment designed for handling samples in porous form.

Disclosure of Invention

The present application describes a lyophilization kit for preparing a lyophilized material comprising: a base comprising a bottom plate having a base edge extending upwardly from a perimeter thereof, the bottom plate having a top surface adapted to support a plurality of reservoirs thereon; a lid comprising a top panel having a lid edge extending downwardly from a perimeter thereof, the top panel having one or more vent holes extending therethrough; an interior space defined by the base and the cover when the base and the cover are in a closed relationship; a gasket located between the base rim and the cover rim and forming a hermetic seal therebetween when compressed; one or more sealing elements each in closeable engagement (e.g., sliding engagement, hinged engagement, swinging engagement, rotational engagement, etc.) with a corresponding one of the vent apertures such that the sealing element and corresponding vent aperture are operable between: (i) an open configuration in which the position of the sealing element relative to the corresponding vent allows fluid communication between the interior space and air outside the cartridge when the base and lid are in a closed relationship; and (ii) a closed configuration in which the position of the sealing element relative to the corresponding vent hole does not allow fluid communication between the interior space and the air outside the cartridge when the base and the cover are in a closed relationship, such that the interior space is sealed from the air outside the cartridge when the base and the cover are in a closed relationship.

In various embodiments, the one or more sealing elements are each a flexible insert comprising a sealing cap and a body portion depending from the sealing cap, the body portion being in closable engagement with a corresponding one of the vent holes and having one or more vent slots formed therein. The sealing elements are each configured to be disposed in a corresponding vent such that (i) when the sealing elements and corresponding vents are in an open configuration, the interior space is in fluid communication with air outside the lyophilization nest via the vent channel; and (ii) the interior space is not in fluid communication with air outside the lyophilization nest when the sealing element and corresponding vent are in a closed configuration such that the bottom surface of the sealing cap is in sealing contact with the top surface of the top plate.

In various embodiments, at least one of the base and the cover comprises aluminum. Various embodiments may include hinges along one or more edges of the top panel or base to provide a clamshell configuration between the base and the cover.

In various embodiments, the base is internally configured to support one or more reservoirs therein. In various embodiments, the reservoirs may each include a plurality of freeze-dried pores configured to be sealed and a plastic and/or low moisture vapor transmission rate material such as a cyclic olefin copolymer. Further, in various embodiments, the top surface of the base plate includes one or more reservoir-receiving portions formed therein, the reservoir-receiving portions each having features that follow at least a bottom end of the one or more reservoirs to be received thereon and configured to conduct heat between the base and the one or more reservoirs supported on the base. In various embodiments, the one or more reservoirs to be supported within the lyophilization nest each comprise a plurality of lyophilization wells, and the reservoir receiving portions each comprise a plurality of well receiving features that follow the shape of at least the bottom ends of the lyophilization wells of the corresponding reservoir. Further, in various embodiments, the lyophilization nest additionally comprises one or more reservoir frames located on the top surface of the floor, the reservoir frames each being configured to support a plurality of reservoirs (e.g., four reservoirs).

In various embodiments, the lyophilization nest further comprises one or more fasteners for securing the lid to the base. The fastener of the lyophilization nest is configured to compress the gasket between the base edge and the lid edge to form an air-tight seal when the base and the lid are in a closed relationship. By way of example, the fastener further includes a base engagement member, such as a base pin, a cover engagement member, such as a cam lever, and a connecting member that couples the other two members. As further examples, the fastener may be a latch, bolt, clamp, or other structure that compresses the gasket between the base and lid edges when the base and lid are in a closed relationship to form an air-tight seal.

Various embodiments of the present application relate to a lyophilization system for lyophilizing one or more substances, the lyophilization system comprising: a sealable housing defining a chamber and having a plurality of shelves. In various embodiments, the plurality of shelves are upper and lower shelves contained within the chamber. In various embodiments, the plurality of shelves is at least one upper shelf and at least one lower shelf. In various embodiments, the plurality of shelves are an upper shelf, a lower shelf, and at least one intermediate shelf, which may be referred to as an upper shelf and/or a lower shelf, depending on its spatial relationship to a lyophilization nest in the lyophilization system. The plurality of shelves are an upper shelf, a lower shelf, and at least two intermediate shelves, which may be referred to as an upper shelf and/or a lower shelf, depending on the spatial relationship of the lyophilization nest in the lyophilization system. In various embodiments, the plurality of shelves are at least one upper shelf and at least one lower shelf, each relative to a lyophilization nest in the lyophilization system. In various embodiments, the lyophilization nest is located on the lower shelf. In various embodiments, at least one of the upper shelf and the lower shelf is capable of automated movement that causes the upper shelf to engage the sealing element, thereby changing the sealing element and corresponding vent from an open configuration to a closed configuration. In various embodiments, the upper shelf is located above the lyophilization nest and is configured to automatically move downward toward a top surface of a lid of the lyophilization nest.

Various embodiments of the present application relate to a lyophilization system for lyophilizing one or more substances, the lyophilization system comprising: a sealable housing defining a chamber and having at least one upper shelf and at least one lower shelf contained within the chamber; and at least one lyophilization nest located on the lower shelf. In various embodiments, at least one of the upper and lower shelves is capable of automated movement that causes the upper shelf to engage a sealing element of a lyophilization nest on the shelf below (the lower shelf) to change the sealing element and corresponding vent from an open configuration to a closed configuration. In various embodiments, the upper shelf is located above the lyophilization nest and is configured to automatically move downward toward a top surface of a lid of the lyophilization nest. Various embodiments relate to methods for lyophilizing one or more substances, comprising: disposing a lyophilization nest on a lower shelf contained within a lyophilizer chamber, wherein the lyophilization nest supports one or more reservoirs contained within an interior space of the lyophilization nest, and at least one of the reservoirs contains one or more substances to be lyophilized. The interior of the closed lyophilization nest is in fluid communication with air outside the nest through one or more vent holes extending through a top plate of a lid of the lyophilization nest, and wherein the vent holes are sealing elements in sliding engagement therewith. The method additionally comprises the steps of: closing the chamber containing the lyophilization nest, creating lyophilization conditions within the chamber for a time sufficient to lyophilize the substance contained in the reservoir. Further, the method may include: moving at least one of a lower shelf and an upper shelf contained within the chamber such that the upper shelf engages the sealing element, thereby closing the vent hole such that the interior space of the lyophilization nest is sealed from air outside the lyophilization nest. The interior space of the sealed lyophilization nest contains environmental conditions from the sealed lyophilization chamber, including low moisture content, and may further include other factors from the environment of the sealed chamber such as nitrogen. The sealed lyophilization nest is no longer in fluid communication with the sealed chamber. The chamber can thus be unsealed so that environmental conditions that are generally not conducive to maintaining the lyophilized substance can be introduced without exposing the lyophilized substance within the sealed lyophilization nest to adverse environmental conditions.

In various embodiments, creating a controlled environment within the lyophilization chamber for lyophilization includes creating a vacuum and cycling between temperatures below freezing that will dry the substance within the chamber. At the end of these steps of the lyophilization process, the humidity level is close to zero. To unseal the lyophilization chamber, the vacuum must first be released. The vacuum is preferably released by flushing the chamber with nitrogen. A high nitrogen environment is preferred due to the low moisture content. The lyophilization nest is preferably sealed after introducing nitrogen gas into the chamber, thereby sealing the interior of the lyophilization nest to a high nitrogen environment. However, this is not mandatory and the lyophilization nest may be sealed at any time after the substances within the lyophilization nest are dried. Once the vacuum has been released, the chamber door can be opened.

In various embodiments, the method may additionally comprise: the reservoirs are placed into and removed from the lyophilization nest, and the reservoirs are sealed, thereby forming an air-tight seal between the wells of the plurality of wells of each reservoir, thereby isolating the interior of the wells from environmental conditions that adversely affect the lyophilized substance in the wells. In various embodiments, the reservoir is sealed as follows: a low moisture vapor permeable membrane (e.g., a laminate structure including an aluminum foil layer) is secured to a top surface of each reservoir to form an air-tight seal between the aperture and the air outside the sealed aperture.

In various embodiments, the relative humidity within the interior space of the lyophilization nest is maintained for an amount of time of at least 4 hours, or at least 8 hours, or from 4 hours to 8 hours at a relative humidity of 10%, more preferably less than 5%, more preferably close to 0%. In various embodiments, the absolute humidity within the interior space of the lyophilization nest is maintained for a period of at least 4 hours, or at least 8 hours, or from 4 hours to 8 hours at 2.3 grams of water per cubic meter of air, more preferably less than 1.15 grams of water per cubic meter of air, more preferably less than 0.23 grams of water per cubic meter of air, more preferably close to 0.0 grams of water per cubic meter of air.

Drawings

Reference will now be made to the accompanying drawings, which are not necessarily drawn to scale, and wherein:

fig. 1 shows a lyophilization nest in a sealed configuration according to one embodiment;

fig. 2 shows an exploded view of a lyophilization nest according to one embodiment;

fig. 3 shows a bottom perspective view of a lid of a lyophilization nest according to one embodiment;

4A-4C illustrate various positions of a sealing element within a vent according to one embodiment;

fig. 5 illustrates a cross-sectional view of a lyophilization nest having a reservoir disposed therein, wherein the upper and lower shelves are in contact with the outer surfaces of the lid and the base, according to one embodiment;

6A-6C illustrate various views of a reservoir according to one embodiment; and

fig. 7-9 include cross-sectional views of a lyophilization nest in various configurations that includes a reservoir in an interior space thereof according to one embodiment. Fig. 7-9 also illustrate chambers including upper and lower shelves shown in various positions outside the cover and base of the kit.

Detailed Description

A lyophilization kit will now be described more fully with reference to the accompanying drawings, in which some, but not all embodiments of the kit are shown. Indeed, the lyophilization nest may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Like reference numerals refer to like elements throughout the specification.

Lyophilization is a well-known method for drying a substance to protect such substance. The main mechanism required for lyophilization is sublimation whereby ice is converted directly to water vapor without passing through an intermediate liquid stage. This is not done by heating, but by removing the pressure so that the ice boils without melting. The result is a substance whose structure remains substantially unchanged, whose water content is negligible, and which can be stored at room temperature and pressure. For the purpose of lyophilizing a substance, many environmental factors surrounding the substance are altered compared to the more typical levels of these factors in the environment. During the lyophilization reaction, many environmental factors frequently change, including one or more of temperature, pressure, atmospheric gas content, and humidity. One skilled in the art of lyophilization will readily develop and implement a set of environmental conditions that will achieve the desired material for lyophilization. The term "controlled environment" and its various forms as used herein refers to an environment in which many of the factors are changed in order to lyophilize a substance or maintain a low water content in a lyophilized substance. As used herein, "uncontrolled environment" and related terms refer to an environment having factors that are detrimental to the lyophilized material or maintain a low water content in the lyophilized material. Note that uncontrolled environments are not used herein to indicate that the environment is not controlled in some way (e.g., room air conditioning, etc.), but are used herein for convenience to indicate environmental conditions that are not conducive to lyophilizing a substance and maintaining a low water content of the lyophilized substance.

Various embodiments relate to a lyophilization kit configured to maintain a controlled environment having at least a low humidity level around one or more substances after a lyophilization process is completed using a lyophilization system. A lyophilization system includes a lyophilizer, a lyophilization nest, and one or more reservoirs each containing one or more substances, the lyophilization system configured to lyophilize the substances and maintain a controlled environment around the substances after a lyophilization process.

Changes in environmental factors are generally within the reach of commercial freeze dryers. Many environmental factors are reduced to negligible levels so that further reductions do not bring significant additional benefits in lyophilizing or preserving samples. One such factor is humidity, where it is desirable to bring the relative water vapor content of air to near zero percent. Humidity can be expressed as a relative value compared to the maximum saturation at a given condition, or as an absolute value measured in grams per cubic meter of water. 1% relative humidity at 25 ℃ equivalent to 0.23g/m³At 25 ℃ and a 5% relative humidity equivalent to 1.15g/m³。

For example, the relative humidity within the lyophilizer can be reduced to about 0% (undetectable by conventional equipment), or less than 1%. The absolute humidity can be reduced to be less than or equal to 0.23g/m³. The controlled environment may also include changing the amount of inert gas (e.g., nitrogen) to a ratio greater or less than the amount present in the ambient atmosphere. The controlled environment may thus be characterized by any or all of the following: reduced atmospheric pressure (vacuum), reduced atmospheric vapor content (humidity), increased inert gas content, or reduced atmospheric temperature as compared to the surrounding uncontrolled environment. Measurements of these changing factors are generally known, as are formulas for converting between their relative and absolute values. Various embodiments of the apparatus and methods of using the same described herein maintain a controlled environment around one or more lyophilized substances after the controlled environment within the lyophilizer has been replaced with an atmosphere that contains factor levels that are less desirable for the lyophilized substances than the controlled environment.

The lyophilization kit is based in part on the following insights: conventional forms for lyophilizing and then resealing the material in the reservoirs (e.g., vials and/or multi-well plates) can result in excessive rehydration of the lyophilized material, possibly due to long contact times with the surrounding uncontrolled environment from the time the seal of the lyophilizer is removed until the individual wells are resealed. Over-hydration can reduce the activity or shelf life of the material or, in some cases, can even lead to exothermic and potentially dangerous reactions. This excessive contact may be caused by a workflow that requires the resealing process to be performed at a workstation located a distance from the lyophilizer, or at a later time after the seal of the lyophilizer is removed. Accordingly, the present application describes apparatus for reducing, minimizing and/or avoiding potentially harmful exposure of a lyophilized substance to undesirable levels of environmental factors in an uncontrolled environment, and thereby absorption of moisture. The apparatus may be used with or without conventional means of reducing sample rehydration, such as glove boxes and drying chambers.

Exposure of the lyophilized substance to undesirable levels of environmental factors in an uncontrolled environment can be avoided by using a sealable lyophilization nest apparatus that preserves the controlled environment formed in the chamber of a sealed lyophilizer inside the apparatus. The device comprises one or more reservoirs in its interior containing one or more substances for lyophilization in its reservoir wells. The interior of the apparatus is in fluid communication with the controlled environment formed within the chamber of the lyophilizer during the lyophilization process and is maintained for a time sufficient to lyophilize the substance. After lyophilization, fluid communication between the lyophilization chamber and the interior of the apparatus is then prevented as follows: the apparatus is sealed so that the interior of the apparatus is isolated from the air in the chamber. The controlled environment within the chamber is then changed as follows: unsealing the chamber and allowing the uncontrolled environment to enter the chamber. However, because the interior of the device is sealed, the environment within the interior of the device is still conducive to maintaining the lyophilized substance therein. It should be noted that during lyophilization, all environmental factors within the apparatus need not be at the same level as the environmental factors of the controlled environment formed within the lyophilization chamber. For example, the temperature inside the device may be different from the temperature used during lyophilization. However, environmental factors inside the sealed device are beneficial for maintaining the lyophilized substance such that rehydration of the lyophilized substance is limited. When the apparatus is not in fluid communication with the environment external to the apparatus, after removal of the seal of the lyophilizer chamber, the apparatus may maintain a favorable environment around and within the reservoir until the operator is ready to retrieve a single reservoir or a well (e.g., a single well) within the reservoir.

A reservoir is a discrete and continuous container for containing one or more substances. The reservoir may have one or more wells, each well for containing a substance. A reservoir having one aperture is referred to as a single-aperture reservoir. A reservoir having a plurality of wells, such as a multi-well plate, is referred to as a multi-well reservoir.

In various embodiments, the device may be sealable and portable such that the device including the reservoir may be removed from the chamber of the lyophilizer and relocated to a suitable workstation (e.g., a clean room, countertop, or drying room) to seal the individual reservoir or individual well therein. The device is referred to in this application as a lyophilization kit, as it provides a temporary storage unit for any lyophilized substance therein for the following periods of time: the time period between removal of the lyophilizer seal and the start of the resealing process of the reservoir containing the lyophilized material.

Briefly, in various embodiments, a substance to be lyophilized is placed in one or more wells of a reservoir. The reservoir is placed within a lyophilization nest configured to support the reservoir within an interior space of the lyophilization nest. The interior space of the lyophilization nest may be in fluid communication with the exterior of the lyophilization nest through one or more vent holes extending through a surface of the lyophilization nest (e.g., a lid of the lyophilization nest) so as to allow air exchange between the interior and the exterior of the lyophilization nest. For example, a lyophilization nest may include a base and a lid configured to collectively define a nest interior space. The lyophilization nest may include a gasket positioned between the base and the cover to form an air-tight seal therebetween when the base and the cover are in a closed relationship. The base may define a bottom panel configured to support one or more reservoirs within the kit interior space, and the lid may define one or more vents through the lid top panel. The vent may have a corresponding sealing element in closeable engagement with the vent, the sealing element being configured to move between an open configuration and a closed configuration. Preferably, the sealing element is configured as a stopper-like insert which is in sliding engagement with the vent and slides between an open configuration and a closed configuration. When the vent of the lyophilization nest is closed, the lyophilization nest is isolated from environmental factors outside the lyophilization nest, thereby preventing air exchange between the inside and outside of the lyophilization nest. The vent may be closed by a mechanism operable from outside the sealed lyophilizer. For example, the sealing element may be pressed into the closed configuration as follows: the shelf within the lyophilizer is lowered against the outer surface of the lyophilization nest cover, thereby engaging and pressing the sealing elements into their respective vent holes.

A lyophilization nest comprising a reservoir is placed within a lyophilizer with one or more vent holes in an open position. The lyophilization chamber is then sealed and the contents of the reservoir wells are lyophilized. During the lyophilization process, atmospheric factors within the lyophilizer change to facilitate the removal of water from the material. The controlled environment within the lyophilization chamber thus enters the interior space of the lyophilization nest by being in fluid communication with the one or more vent holes. The one or more vent holes are then closed, thereby sealing the lyophilization nest to prevent rehydration of the substance after the lyophilization process is complete by maintaining environmental conditions within the lyophilization nest that are favorable for maintaining the lyophilized substance. The seal on the lyophilizer is later removed and the lyophilization nest containing the reservoir is removed from the chamber. The lyophilization nest can then be relocated and stored with the sample container in the nest until the operator is ready to use the lyophilized material therein or reseals the reservoir containing the lyophilized material for further storage or sale. Various embodiments of the lyophilization nest may be used with a lyophilizer having a vertically movable shelf that may be repositioned when the lyophilizer is sealed. Thus, the vertically movable shelf may be lowered against the lyophilization nest to engage the sealing element, thereby sealing the vent of the lyophilization nest while maintaining a controlled environment within the lyophilizer.

The components of the lyophilization nest, i.e., the reservoirs, seals, and the like, may be made using materials that include a low moisture vapor transmission rate. Moisture vapor transmission rate is a measure of the passage of water vapor through a substance. There are many methods for determining the moisture vapor transmission rate of a material (e.g., many standard methods are described by the international organization for standardization (ISO), the American Society for Testing and Materials (ASTM), etc.). Additionally, materials having reported moisture vapor transmission values are commercially available. One of ordinary skill in the art will know how to calculate and/or purchase a material having a low moisture vapor transmission rate.

Freeze-drying kit

Referring now to fig. 1, which illustrates an exemplary lyophilization nest 1 in a closed configuration, lyophilization nest 1 may define an airtight container configured to maintain a controlled environment therein. As shown in the embodiment of fig. 1, the lyophilization nest 1 may comprise a base 10 and a cover 50, with a gasket 60 positioned between the base 10 and the cover 50, forming an air-tight seal between the base 10 and the cover 50 when the gasket 60 is compressed between the base 10 and the cover 50. Referring briefly to fig. 2, lid 50 defines one or more vent holes 53 in top plate 52, which vent holes 53 are configured to selectively place the interior space of lyophilization nest 1 in fluid communication with the atmosphere surrounding lyophilization nest 1. As shown in the embodiment of fig. 1, the one or more vent holes 53 each have a corresponding sealing element 100, such as a rubber insert or other flexible insert. In fig. 1, the sealing elements 100 are each slidably engaged with a corresponding vent hole 53, the vent holes 53 being configured to engage with a perimeter of the vent holes 53, thereby forming an airtight seal therein.

Further, in the illustrative embodiment of fig. 1, the lyophilization nest 1 comprises one or more fasteners 150 (specifically latches), the one or more fasteners 150 configured to selectively secure the base 10 and the cover 50 and compress the gasket 60 between the base 10 and the cover 50 to form an airtight seal therebetween. However, various fasteners may be used to secure the base 10 and cover 50 (e.g., clamps, screws, bolts, nuts, etc.). Further, in various embodiments, the base 10 and cover 50 may be secured via one or more hinges configured such that the base and cover may be rotated about a hinge axis between an open relationship and a closed relationship. In such an arrangement, one or more fasteners may be used in conjunction with one or more flaps such that the base 10 and lid 50 may be sealed in a closed relationship to form an airtight seal therebetween.

In various embodiments, multiple bases and/or multiple lids may collectively define a lyophilization nest 1. As a non-limiting example, two or more lids 50 may be configured to be secured with a single base 10 in a closed configuration to define a lyophilization nest 1. When each of the plurality of covers 50 is engaged with the base 10 in the closed configuration, the plurality of covers may be configured to form an airtight seal therebetween. For example, one or more gaskets may be located between each of the plurality of lids to form an air-tight seal therebetween when the lyophilization nest is in the closed configuration. As an example, two covers 50 may each be secured to the other side of a single base 10 via one or more hinges. Thus, the two covers may be configured to rotate between an open configuration in which the covers do not prevent access to the interior of the base 10 and a closed configuration in which an airtight seal is formed between the two covers and the base 10.

In various embodiments, the base 10 comprises a metallic material (e.g., aluminum) and the cover 50 comprises a plastic material (e.g., a high density polyethylene material, a polyvinyl chloride material, etc.), although other materials are also contemplated. For example, the base 10 and the cover 50 may each comprise aluminum and/or a plastic material. The base 10 and the cover 50 are each of a material that at least substantially prevents moisture from migrating into the interior space of the lyophilization nest 1. Thus, for example, any material having a low moisture vapor transmission rate may be used (e.g., cyclic olefin copolymer materials; Topas Advanced Polymers, Inc., Florence, KY; catalog number 8007S-04). As another example, other materials may be used in amounts that provide a wall thickness of the base 10 and/or the cover 50 so as to prevent vapor from passing through the base and cover between the interior and exterior of the lyophilization nest 1. By way of non-limiting example, the base 10 and/or the cover 50 may each comprise a metal material, a plastic material, a composite material, or the like.

Fig. 2 shows an exploded view of the various components of lyophilization nest 1, providing additional details of each of the various components of lyophilization nest 1. In the illustrative embodiment of fig. 2, the base 10 includes a raised base rim 11 surrounding a bottom panel 12, the base rim 11 and a top surface of the bottom panel 12 together defining a base interior 15. As shown in fig. 1 and 2, the base 10 may be at least substantially rectangular, but the base 10 may have any of a variety of shapes (e.g., circular, triangular, etc.). In various embodiments, the raised base edge 11 includes a smooth sealing surface (not shown in fig. 2) configured to engage the gasket 60 to form a hermetic seal therebetween when the gasket 60 is compressed between the raised base edge 11 and a corresponding lid edge, as discussed herein.

In the illustrative embodiment of fig. 2, the base interior 15 is configured to support a plurality of individual reservoirs 200 therein. As will be discussed in greater detail herein, the plurality of reservoirs 200 may each define one or more substance wells, each substance well configured to hold a substance to be lyophilized. For example, the reservoirs 200 may each be a single-pore reservoir or a multi-pore reservoir. The base interior 15 may include one or more reservoir receiving portions 16 therein. Each of the one or more reservoir-receiving portions 16 may be integrated into the base 10. In various embodiments, one or more receiving portions 16 may be secured to the base 10. As with the base 10, the one or more reservoir-receiving portions 16 may comprise aluminum; but a variety of alternative materials are also contemplated. In various embodiments, the one or more reservoir-receiving portions 16 may be configured to conduct heat away from the one or more reservoirs 200 located therein, and/or may be configured to support the one or more reservoirs 200 in an upright position. Thus, the one or more reservoir-receiving portions 16 may be configured to support the one or more reservoirs 200 to be disposed therein and/or follow the external shape of at least the bottom of the one or more reservoirs 200 to be disposed therein to maximize the surface contact area between the reservoirs 200 and the corresponding reservoir-receiving portions 16. Further, the one or more reservoir-receiving portions 16 may comprise a material having a high heat transfer coefficient (e.g., aluminum) and may be in sufficient contact with the base 10 such that heat transfer between the one or more reservoir-receiving portions 16 and the base 10 is maximized. Referring again to the exemplary lyophilization nest 1 shown in fig. 2, the one or more reservoir-receiving portions 16 are formed of a material that is a unitary piece of material with the base 10. In various embodiments, one or more reservoir receiving portions 16 may be removably secured to the base 10.

In the illustrative embodiment of fig. 2, the lyophilization nest 1 additionally comprises a lid 50 having a shape corresponding to the base 10. In the illustrated embodiment, the cover 50 has a rectangular shape corresponding to the base 10. Alternative shapes (e.g., circular, triangular, etc.) are also contemplated.

As shown in fig. 3, which illustrates a bottom view of lid 50 according to various embodiments, lid 50 has a lid edge 51 that surrounds the perimeter of top panel 52 and extends away from top panel 52 to define a lid interior 55. In an exemplary embodiment, the lid edge 51 includes an at least substantially smooth surface configured to form an airtight seal with a gasket 60 (not shown in fig. 3) and a corresponding base edge 11 (fig. 2) when the gasket 60 is compressed between the lid edge 51 and the base edge 11. Further, in various embodiments, the gasket 60 may be secured to one of the lid 50 or the base 10 (fig. 2) such that the sealing portion of the gasket 60 is located between the base edge 11 and the lid edge 51.

Although the illustrative embodiment of fig. 1-3 shows base rim 11 and lid rim 51 having substantially planar surfaces configured to compress gasket 60 between base rim 11 and lid rim 51 to provide an airtight seal, various embodiments may include other configurations for providing an airtight seal between the components of lyophilization nest 1. As a non-limiting example, the base rim 11 and the lid rim 51 may have an interlocking configuration such that the surfaces of the base rim 11 and the lid rim 51 may include corresponding features such that the base rim 11 and the lid rim 51 may interlock when the lyophilization nest 1 is in the closed configuration. For example, the base rim 11 and the lid rim 51 may have a tongue and groove arrangement such that at least one of the base rim 11 and the lid rim 51 may include a tab configured to engage a corresponding groove on the other to form a seal therebetween. In various embodiments, a gasket may be disposed between the base rim 11 and the lid rim 51 (e.g., within a groove and/or on a tab).

As shown in fig. 2 and 3, lid 50 may define one or more vent holes 53, which vent holes 53 extend through top plate 52 from the top surface of top plate 52 to the inner surface of top plate 52, such that the interior volume of lid interior 55 is in fluid communication with the air surrounding lyophilization nest 1 through vent holes 53. The fluid communication through the vent 53 allows air exchange between the interior space of the lyophilization nest 1 and the exterior of the lyophilization nest 1. In various embodiments, the one or more vent holes 53 may each correspond to a separate compartment within the lyophilization nest 1 that is separate from other separate compartments when the lid 50 is secured to the base 10. However, in various embodiments, one or more vent holes 53 may each place a single interior space of the lyophilization nest in fluid communication with air surrounding the lyophilization nest 1. As shown in fig. 2, each vent hole 53 may have a corresponding sealing element 100 (e.g., a gasket, rubber insert, or other flexible insert) slidably engaged with the corresponding vent hole. Sealing element 100 is configured to engage the perimeter of vent hole 53, thereby forming an air-tight seal within vent hole 53, which isolates the interior space of lyophilization nest 1 (collectively defined by base interior 15 and lid interior 55) from air surrounding the exterior of lyophilization nest 1 (exterior of base 10 and lid 50). As shown in fig. 2, at least a portion of the sealing element 100 may be located outside of the sealable lyophilization nest 1 and may be configured to be at least partially pressed into the top plate 52 to form an airtight seal in the corresponding vent hole 53. Further, as shown in fig. 1 and 2, the top plate 52 may define a recess 54 around each of the one or more vent holes 53. In the illustrative embodiment of fig. 1 and 2, the recess 54 may have a uniform depth throughout the recess 54, thereby defining a recess surface on the bottom of the recess 54, and may be configured such that a portion of the sealing element 100 rests against the recess surface when sealed within the corresponding vent hole 53. Further, the recesses 54 may have a depth such that a top surface of the sealing member 100 is at least substantially aligned with an outer surface of the top plate 52 when the sealing member 100 is sealed within the corresponding vent hole 53.

Although the illustrative embodiment of fig. 1-3 shows one or more vent holes 53 extending through the top plate 52 of the lid, various freeze-drying kit 1 embodiments have vent holes 53 extending through other portions of the freeze-drying kit 1. For example, one or more vent holes 53 may extend through a side of lyophilization nest 1, and/or through base 10. In various embodiments, a lyophilizer may include a lyophilization nest sealing system configured to engage and move one or more sealing elements to a closed configuration. In various embodiments, the lyophilization nest sealing system may be configured to operate when sealing the lyophilizer.

Fig. 4A-4C illustrate various positions of the sealing element 100 relative to the corresponding vent 53 that are operable between an open configuration (fig. 4A and 4B) and a closed configuration (fig. 4C). Fig. 4A-4C illustrate an optional recess 54 in which is a vent 53. As shown in fig. 4A and 4B, each sealing element 100 may define a sealing cap 101, a body portion 102, and a vent channel 103, the vent channel 103 being configured such that when the corresponding sealing element 100 is loosely disposed in the one or more vent holes 53, the interior space of the lyophilization nest 1 may be maintained in fluid communication with air surrounding the lyophilization nest 1 through the one or more vent holes 53. In various embodiments, the sealing cap 101 may define a top surface of the sealing element 100. As shown in fig. 4A-4C, the sealing cap 101 is illustrated as a circular element having a flat top surface and a flat bottom surface. The sealing cap 101 is secured to the body portion 102 of the sealing element 100. As shown in fig. 4A-4B, a top portion of the body portion 102 of the sealing element is secured to a bottom surface of the sealing cap 101. In various embodiments, the edge of the sealing cap 101 may extend beyond the perimeter of the body portion 102 such that a portion of the bottom surface of the sealing cap 101 is configured to be in sealing contact with the top surface of the top plate 52 when the sealing element 100 is inserted into the corresponding vent hole 53 in the closed configuration. Referring to fig. 4A-4B, as a specific example, the body portion 102 and the sealing cap 101 of the sealing element 100 may be concentric circles, wherein the diameter of the sealing cap 101 is greater than the diameter of the body portion 102.

4A-4B, the sealing element 100 may have a vent slot 103 extending through at least a portion of the body portion 102. For example, as shown in fig. 4A and 4B, the vent slot 103 may extend from a bottom portion of the body portion 102 to a top portion of the body portion 102 and terminate between the bottom portion and the top portion of the body portion 102. In various embodiments, the vent slot 103 may extend across the entire diameter of the body portion 102. Further, in various embodiments, the sealing element 100 may comprise a one-piece flexible material. Thus, in the situation as shown in fig. 4B, i.e. with the sealing element 100 in an open configuration, wherein the sealing element 100 is loosely placed within the corresponding vent hole 53, such that a portion of the vent channel 103 is located outside the top surface of the lid 50, the inner space of the lyophilization nest 1 remains in fluid communication with the air surrounding the outside of the lyophilization nest 1. When one or more sealing elements 100 are pressed into the corresponding vent 53 (fig. 4C) in the closed configuration such that the bottom surface of the sealing cap 101 is in sealing contact with the top surface of the lid 50, the interior space of the lyophilization nest 1 is sealed off from the exterior of the lyophilization nest and the interior space of the lyophilization nest is no longer in fluid communication with the exterior of the lyophilization nest. In various embodiments, one or more sealing elements 100 may comprise a rubber material, although a variety of resilient and/or flexible materials (preferably having a low vapor transmission rate) may also be used. Furthermore, one or more sealing members 100 may be configured to operate when lyophilization nest 1 is located within a lyophilizer. For example, as will be described in greater detail in the present application, one or more sealing elements 100 may be configured to be pressed into a sealed position within a corresponding vent hole 53 as follows: a vertically movable shelf or other automatically movable actuating element is moved within the lyophilizer towards the outer surface of the lyophilization nest 1 (e.g. downwardly towards the lid 50) to engage one or more sealing elements 100, thereby changing the sealing elements and corresponding vent holes 53 from an open configuration to a closed configuration.

Fig. 5 illustrates a cross-sectional view of the lyophilization nest 1 in a closed and sealed configuration, between the upper shelf 301 and the lower shelf 302 within the chamber 401 of the lyophilizer. As shown in fig. 5, when the cover 50 is positioned such that the cover edge 51 is aligned with the base edge 11, the base interior 15 and the cover interior 55 form an interior space of the lyophilization nest 1. In the illustrative embodiment of fig. 1, 2 and 5, lyophilization nest 1 additionally comprises one or more fasteners 150 configured to secure lid 50 relative to base 10 and thereby maintain an airtight seal therebetween. As illustrated in fig. 2, the one or more fasteners 150 each include a base pin 151, a cam lever 152, and a connecting arm 153, although multiple fasteners may be used to secure the cover 50 to the base 10. The fastener is configured to selectively compress the gasket 60 between the cover 50 and the base 10 by pressing the base 10 toward the cover 50.

Further, as shown in fig. 5, the one or more reservoir receiving portions 16 include features for supporting and/or following at least the bottom end of the reservoir 200 to be received therein. In the illustrative example of fig. 5, the reservoir-receiving portions 16 each include a plurality of reservoir aperture-receiving features having a shape that follows the external shape of an aperture 201 formed within the reservoir 200. For example, for a reservoir aperture 201 having a hemispherical exterior, the reservoir aperture receiving feature may have a corresponding hemispherical shape. Such a corresponding shape may maximize the surface contact area of each of the one or more apertures 201 in contact with the reservoir-receiving portion 16 to maximize conductive heat transfer between the substance located within each of the one or more apertures 201 and the base 10.

In the exemplary embodiment of fig. 2, the lyophilization nest 1 is additionally configured to support one or more reservoir frames 250, the reservoir frames 250 each having one or more reservoir support portions 251. As shown in fig. 2, the one or more reservoir frames 250 are each configured to support the one or more reservoirs 200 therein, thereby facilitating placement and removal of the one or more reservoirs 200 within the lyophilization nest 1. As shown in the illustrative embodiment of fig. 2, the one or more reservoir frames 250 may be configured to be positioned on the top surface of the bottom panel 12 and around the one or more reservoir receiving portions 16 and extend above the top surface of the base rim 11 when positioned within the base 10. Thus, the perimeter of the one or more reservoir frames 250 may thereby provide a positioning guide for ensuring proper positioning of the lid 50 relative to the base 10 such that an airtight seal may be formed therebetween. However, in various embodiments, at least one of the cover 50 and/or the base 10 may include one or more extrusions (extrusions) that extend beyond the surface of the

corresponding edges

11 and 51 to provide a locating member for positioning the cover 50 relative to the base 10. Further, in various embodiments, the one or more reservoir frames 250 may each comprise aluminum, although any of a variety of rigid materials (e.g., plastics, other metals, composites, etc.) may also be used.

Fig. 6A-6C illustrate a reservoir 200 that can be disposed within a lyophilization nest 1 according to various embodiments. As shown in fig. 6A and 6C, the one or more reservoirs 200 can each define one or more apertures 201 configured to receive a substance to be lyophilized. For example, the one or more reservoirs 200 may each include a single aperture or a plurality of apertures. The one or more reservoirs 200 can each comprise a low vapor permeability material (e.g., cyclic olefin copolymer) configured to prevent water vapor migration through the material, although other materials (e.g., polypropylene) can also be used. Further, the one or more reservoirs may each have a sealable upper surface 202, said upper surface 202 being configured such that a seal (e.g., an adhesive low moisture vapor permeable material secured to the sealable upper surface 202 of the reservoir 200) may be applied thereto, said upper surface 202 being sealed such that the one or more wells are each individually sealable and isolated from the uncontrolled environment outside the sealed reservoir and/or wells. For example, as shown in fig. 6B, a seal 203 may be secured and sealed to the top of the reservoir 200 and around each of the one or more apertures 201 (not shown in this figure). In a preferred embodiment, the seal 203 is a low moisture vapor transmission film, such as a laminate structure including an aluminum foil layer, but the seal may be made of other materials. One or more apertures 201 may each be individually accessed to gain access to the aperture as follows: the seal 203 is pierced, the seal 203 is peeled away from the sealable upper surface 202, or otherwise removed from over one of the apertures 201. Removal of the seal 203 (e.g., puncturing) to provide access to the lyophilized substance in the underlying aperture 201 forces the aperture to be in fluid communication with the air outside the reservoir 200. The lyophilized substance in the well 201 from which the seal 203 has been removed may then be purposefully rehydrated or otherwise manipulated, depending on the desired use of the reconstituted agent. In a sealed multi-well reservoir 200, the lyophilized substance of the other wells of the one or more wells 201 in which the seal 203 described above has not been pierced or removed can be stored for later use. As a further example, one or more apertures 201 may each have an associated sealing cap configured to be secured within the aperture to provide a hermetic seal around the perimeter of the aperture.

As shown in fig. 6C, fig. 6C is a cross-sectional view of an array of apertures 201 of a reservoir 200 according to various embodiments, the plurality of apertures may each have an at least substantially hemispherical bottom interior surface. Further, as shown in fig. 6C, the exterior of each of the plurality of apertures 201 may have a corresponding at least substantially hemispherical surface. As described herein, at least the bottom end of the outer surface of each of the plurality of wells 201 can be configured to follow the reservoir well receiving features of the reservoir receiving portion 16 of the lyophilization nest 1. As previously described, such corresponding features may maximize surface area contact between aperture 201 and reservoir-receiving portion 16, thereby facilitating the transfer of heat away from the substance located within aperture 201.

Substances that can be lyophilized by the device and methods of use thereof include, for example, pure chemicals, chemical mixtures, biological samples such as cells, cell extracts, or tissues, biological agents such as nucleic acids, enzymes, antibodies, and other proteins, markers such as fluorophores, and combinations thereof. Other examples of substances that can be lyophilized also include various reaction mixtures for carrying out the following specific reactions: for example, for performing Polymerase Chain Reaction (PCR), transcription mediated amplification, nucleic acid or protein capture assays, and nucleic acid or protein hybridization assays.

By porous reservoir is meant a continuous container that may contain at least two substances such that they may be stored and handled in parallel but separately. Standard formats for multi-well reservoirs include 6 wells, 24 wells, 96 wells, 384 wells, or 1536 wells. The volume of each well in the example 96 well format was about 300-400. mu.L, with a working volume of about 75-200. mu.L. The volume is generally inversely proportional to the number of pores, and is generally in the range of 1nL to 10mL for each pore volume, although other sizes are contemplated. Exemplary holes may have a flat bottom, a hemispherical bottom, or a V-shaped bottom, among others.

As used herein, an example lyophilizer comprises a sealable housing defining a chamber 401, said chamber 401 being configured to support one or more sample containers (e.g., porous sample containers) and/or one or more lyophilization nest(s) 1 therein. The lyophilizer is configured to create and maintain a controlled environment having the conditions necessary to lyophilize one or more substances within a sample reservoir located within chamber 401. An exemplary lyophilizer is configured to adjust the levels of various atmospheric factors, such as air pressure/vacuum, temperature, moisture content and gas content. Preferably, the freeze dryer regulates atmospheric pressure factors as follows: the pressure within chamber 401 is reduced, thereby creating a vacuum within chamber 401. By way of non-limiting example, the pressure within chamber 401 of the lyophilizer can be reduced to 730 mTorr or less, more preferably to 65 mTorr or less, or increased to greater than 760 mTorr. The lyophilizer also preferably removes heat from the chamber 401, thereby lowering the temperature within the chamber 401. Thus, the lyophilizer is configured to create an atmospheric temperature within the chamber 401, any water within the substance in the chamber 401 will freeze and will sublimate out of the substance in view of the air pressure. The lyophilizer also preferably varies the amount of gas in chamber 401, for example by increasing the amount of nitrogen in chamber 401. For example, after the lyophilization process is complete (e.g., after substantially all of the water is removed from the substance), chamber 401 may be filled with an inert displacement gas (e.g., pure dry nitrogen gas), thereby raising the pressure within chamber 401 to at least substantially atmospheric pressure. For example, pure dry nitrogen gas may be introduced into chamber 401 of a lyophilizer while maintaining the pressure within chamber 401 at less than atmospheric pressure (e.g., 730 mtorr or less). However, in various embodiments, the lyophilizer may not introduce an inert gas into chamber 401, thereby maintaining a vacuum within chamber 401 of the lyophilizer at a pressure of less than 730 mtorr, preferably less than 65 mtorr. In still other embodiments, an inert gas may be introduced into chamber 401 of the lyophilizer to raise the pressure within chamber 401 to atmospheric pressure (e.g., 760 mtorr) or higher. The humidity of the resulting controlled environment (e.g., by sublimation followed by introduction of an inert gas) is negligible, which facilitates lyophilization of the material and prevents rehydration of the lyophilized material after lyophilization. Such negligible moisture levels may be characterized in that further reduction of moisture does not provide a significant additional benefit in preventing rehydration of the one or more lyophilized substances. The levels of various environmental factors are adjusted and controlled according to the substance to be lyophilized, thereby lyophilizing the substance. One skilled in the art of lyophilization will readily develop and implement a controlled set of environmental conditions to achieve lyophilization of a useful substance.

The lyophilizer may be adapted to include one or more vertically movable shelves and/or other actuating elements that may be repositioned using a user control system when sealing chamber 401. Such vertically movable shelves and/or other actuating elements may be used to apply pressure to one or more sealing elements, thereby pressing the sealing elements into respective vent holes in the lyophilization nest. In various embodiments, a lyophilizer may include a lyophilization nest sealing system configured to engage and move one or more sealing elements of a lyophilization nest to a closed configuration. For example, a lyophilization nest sealing system may include a sealing member configured to engage one or more sealing elements and move the sealing elements to a closed configuration. The freeze dryer and the freeze drying kit 1 together define a freeze drying system.

Application method

Referring to fig. 7-9, illustrating the various steps for sealing the lyophilization nest 1, the present application describes a method of maintaining a controlled environment around one or more substances using the lyophilization nest 1.

According to various embodiments, one or more substances to be lyophilized are placed within the reservoir 200. For example, each of a plurality of separate substances is placed within a single well of the porous reservoir 200. Although the reservoir 200 is illustrated as separate from each lyophilization nest 1 assembly, in various embodiments, one or more reservoirs may be integrated into one of the plurality of lyophilization nest assemblies (e.g., the base 10). One or more reservoirs 200 are disposed on the top surface of the base plate 12 and within the base interior 15. In various embodiments, one or more reservoirs 200 are each disposed within the base interior 15 relative to a corresponding reservoir receiving portion 16. For example, for a reservoir 200 having one or more individual apertures, the corresponding reservoir-receiving portion 16 may comprise a single aperture-receiving portion having an inner surface corresponding to the outer shape of each of the one or more individual apertures of the reservoir 200. Further, in various embodiments, the one or more reservoirs 200 are each disposed within the reservoir frame 250 prior to being disposed within the base interior 15. For example, as shown in fig. 2, four (4) individual reservoirs 200 may be disposed within a single reservoir frame 250, and then the reservoir frame 250 may be placed in the base interior 15.

After the one or more reservoirs 200 containing the one or more substances to be lyophilized are placed within the base interior 15, the lid 50 is placed on the base 10 such that the base interior 15 and the lid interior 55 collectively form a kit interior space with the one or more reservoirs 200 disposed therein. In various embodiments, one or more fasteners 150 (e.g., latches, not shown) engage the base 10 and the lid 50 to compress the gasket 60 between the base edge 11 and the lid edge 51 to form an airtight seal therebetween when the base 10 and the lid 50 are in a closed relationship. However, in various embodiments, the lid 50 is loosely placed on the base 10 such that an air-tight seal may be formed when the lyophilization nest 1 is disposed within the chamber 401 of the lyophilizer.

As shown in fig. 7, one or more sealing elements 100 (e.g., rubber inserts) are loosely placed within corresponding vent holes 53 of the lid 50 such that the interior space of the lyophilization nest 1 is maintained in fluid communication with the ambient conditions surrounding the exterior of the lyophilization nest (e.g., conditions within the chamber 401 of the lyophilizer) via the one or more vent holes 53. As described herein, by loosely placing one or more sealing members 100 within the corresponding vent 53 such that the sealing members 100 are in an open configuration and the interior space of the lyophilization nest 1 is in fluid communication with the exterior of the lyophilization nest 1 via the one or more vent 53, the sealing members 100 are slidably engaged with the corresponding vent 53 such that the sealing members 100 can be pressed into a closed configuration to seal the vent 53.

The assembled lyophilization nest 1 is then placed within a lyophilizer having a sealable housing defining a chamber 401, the chamber 401 having a seal configured to maintain an environment therein for lyophilizing one or more substances and having at least one vertically movable shelf configured to be automatically moved and manipulated from outside the lyophilizer while the chamber 401 is sealed. For example, the lyophilization nest 1 is placed within the chamber 401 on the lower shelf 302 below the at least one upper shelf 301. A controlled environment is formed within the chamber 401. The interior space of lyophilization nest 1 is in fluid communication with chamber 401, and thus the one or more substances are each in contact with the controlled environment and are lyophilized. After lyophilization of the one or more substances while maintaining the controlled environment within the chamber 401, each of the one or more vent holes 53 is sealed by pressing the corresponding sealing member 100 into the vent hole 53 of the top plate 52 of the lid 50. One or more factors of the controlled environment within the chamber 401 are then maintained in the interior space of the lyophilization nest 1 while the chamber 401 is unsealed, such that the environment in the chamber changes to a level that includes environmental factors that are detrimental to the lyophilized substance. Fig. 8-9 illustrate one embodiment for sealing the lyophilization nest 1 in chamber 401 such that the controlled environment in chamber 401 may be replaced with ambient air while maintaining the controlled environment within the sealed lyophilization nest 1.

As shown in fig. 8-9, the upper shelf 301 within the chamber 401 is moved to engage one or more sealing members 100 and contact the outer surface of the top plate 52 of the lid 50 such that each sealing member 100 is pressed into a corresponding vent hole 53 to form an airtight seal, thereby closing the vent holes 53 to seal the interior space of the lyophilization nest 1. As a non-limiting example, the vertically movable upper shelf 301 may be lowered onto the outer surface of the top plate 52 of the lid 50 while the sealing elements 100 are loosely seated within the corresponding vent holes, allowing the lyophilizer chamber to be in fluid communication with the interior space of the lyophilization nest via the exposed vent slots 103 and vent holes 53 (fig. 7 and 8). The upper shelf 301 is further lowered vertically onto the outer surface of the top plate 52 of the lid 50 and the sealing member is completely pressed into the corresponding vent hole 53, as shown in fig. 9. As another non-limiting example, the lower shelf 302 on which the lyophilization nest 1 is disposed may be moved upward such that the outer surface of the lyophilization nest 1 contacts a surface located above the lyophilization nest 1 such that the sealing elements are pressed into the corresponding vent holes 53. As another non-limiting example, a lyophilization nest sealing system may engage one or more sealing elements 100 and move the sealing elements into a closed configuration. Referring again to fig. 4B-4C, the sealing member 100 may be pressed into the corresponding vent hole 53 from the relaxed position shown in fig. 4B to the sealed position shown in fig. 4C, wherein a portion of the sealing member 100 is in contact with the bottom recess surface of the corresponding recess 54 and the top surface of the sealing member 100 is at least substantially aligned with the outer surface of the top plate 52. Particularly when the one or more sealing members 100 are sealed within the corresponding vent holes 53 while a vacuum is present within the chamber 401 of the lyophilizer, depressing the one or more sealing members 100 until the top surface of the one or more sealing members 100 is substantially aligned with the outer surface of the top plate 52 may prevent the one or more sealing members 100 from being secured to the smooth surface of the upper shelf 301.

In various embodiments, an airtight seal may be formed between the lid 50 and the base 10 when the lid 50 and the base 10 are in a closed relationship as follows: the cover 50 is pressed against the base 10 thereby compressing the gasket 60 therebetween. As a non-limiting example, when a vacuum is formed within chamber 401 of a lyophilizer, an air tight seal may be formed between base 10 and lid 50 while lyophilization nest 1 is located within chamber 401. After releasing the vacuum within chamber 401, the negative pressure within the interior space of lyophilization nest 1 may maintain an airtight seal between base 10 and lid 50.

Once the interior space of lyophilization nest 1 is isolated from the ambient environment within chamber 401 by the hermetic seal within each of the plurality of vent holes 53 and between cover 50 and base 10, the environment within chamber 401 of the lyophilizer can be replaced with air that is not conducive to lyophilization (e.g., high moisture content) as follows: causing air outside of lyophilization chamber 401 to rush into chamber 401. For example, an access door and/or vent of chamber 401 may be opened, thereby replacing the environment within chamber 401 with the environment from the space outside the lyophilizer. Thus, the gas composition, gas pressure, temperature, and/or humidity level within chamber 401 may be altered to be equivalent to the environment surrounding the lyophilizer (e.g., the environment of the chamber in which the lyophilizer is placed). Since the interior space of the lyophilization nest 1 is isolated from the surrounding environment, a controlled environment within the lyophilization nest 1 is maintained.

By way of non-limiting example, a vacuum may be maintained within lyophilization nest 1 for at least 4 hours. As another non-limiting example, in various embodiments, a nitrogen-filled environment may be maintained within lyophilization nest 1 for at least 8 hours. The absolute humidity in the interior space of the lyophilization nest 1 can be less than or equal to 0.23g/m³Maintained for at least 4 hours.

Preferably, the lyophilized material in the lyophilization nest 1 is sealed directly in the reservoir 200 for later use, which requires opening the lyophilization chamber 401, removing the sealed lyophilization nest 1 therefrom and placing it in a processing station, and sealing the reservoir 200. The environmental conditions inside the lyophilization nest 1 facilitate maintaining the lyophilized substance during the process of removing the lyophilization nest 1 from the chamber 401 and sending it to the processing station, from the time the chamber 401 is opened, until the time immediately before unsealing the lyophilization nest 1 to access the reservoirs 200 therein. To access the one or more reservoirs 200 and the lyophilized material stored therein, the lid 50 is removed from the base 10, the reservoirs 200 are removed from the lyophilization nest 1 and sealed for storage and later use or other additional processing. Thus, the exposure time of the lyophilized substance within the lyophilization nest 1 to an uncontrolled environment is reduced, thereby preventing water absorption and thereby maintaining the integrity of the lyophilized substance.

When lyophilizing a bulk material in a single lyophilization process, multiple lyophilization nest 1 are used to maintain a controlled environment inside each lyophilization nest 1 and around the lyophilized material therein, as described herein. To accommodate a plurality of lyophilization nest 1 in the chamber 401, the chamber may include three or more shelves. In configurations where chamber 401 comprises three or more shelves, there is one true upper shelf 301, one true lower shelf 302, and at least one intermediate shelf. In this configuration, each intermediate shelf may serve as both an upper shelf 301, which presses the sealing element 100 onto the top surface of the underlying lyophilization nest 1 when acting as an upper shelf, and a lower shelf 302, on which the lyophilization nest 1 rests when acting as a lower shelf. Here, a plurality of lyophilization nest 1 located on a plurality of the three or more shelves may seal between the three or more shelves within chamber 401 in a concertina-type action. Other configurations may also be used to use multiple lyophilization kits 1 in chamber 401. Once the plurality of lyophilization nest 1 are sealed, the lyophilizer is opened, allowing air to flow into chamber 401. A controlled environment is maintained in the inner space of the sealed lyophilization nest 1 while these lyophilization nests 1 are transferred to a station for further processing of one or more reservoirs 200 therein. A subset (e.g., 1 or less than all) of the plurality of sealed lyophilization kits 1 are then unsealed to process the reservoirs 200 therein. For example, one or more of the plurality of lyophilization nest 1 are unsealed by removing fasteners 150 and separating lid 50 from base 10. The reservoir 200 is covered with a seal 203 for storage and later use. Additional subsets of the plurality of lyophilized kits 1 are then unsealed and subsequently processed. In this example, a plurality of sealed lyophilization nest 1 for lyophilizing a bulk substance maintains a controlled environment around the lyophilized substance in the interior space of each of the plurality of lyophilization nest 1 for the following periods of time: during the introduction of adverse environmental factors into the chamber 401 of the freeze dryer, during the transport of the plurality of sealed freeze-dried kits 1 from the freeze dryer to a subsequent processing station, and while continuing with the subsequent processing.

As mentioned, after removing the seal on the lyophilization nest 1, the lyophilized substance may be used as is, or the reservoir 200 may be sealed for storage and later use of one or more lyophilized substances contained in the one or more wells 201 of the reservoir. Resealing does not remove the ambient air present in the one or more apertures 201 of the reservoir 200, but such a process isolates the substance from prolonged contact with adverse environmental factors and thereby greatly improves the storage of the lyophilized substance. The reservoir 200 may be sealed as follows: the reservoir is removed from the lyophilization nest 1 and a seal 203 (e.g., a low moisture vapor permeable membrane, such as a laminate comprising a layer of aluminum foil) is applied and affixed to the open face of the one or more apertures 201 to form an air-tight seal between each aperture of the reservoir and the outside ambient air. The reservoir 200 comprising more than one aperture is preferably sealed with a seal 203 such that each aperture is individually closed and such that the seal on each aperture can be broken independently without exposing the remaining apertures to adverse environmental conditions.

Conclusion

Many modifications and other embodiments of the lyophilization kits set forth herein will come to mind to one skilled in the art to which these devices and methods of use pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the lyophilization kit and method of using the same are not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

In summary, the present invention includes but is not limited to:

1. a lyophilization kit for preparing a lyophilized material, the kit comprising:

a base comprising a bottom plate having a base edge extending upwardly from a perimeter thereof, the bottom plate having a top surface adapted to support a plurality of reservoirs thereon;

a lid comprising a top panel having a lid edge extending downwardly from a perimeter thereof, the top panel having one or more vent holes extending therethrough;

an interior space defined by the base and the lid when the base and the lid are in a closed relationship, the interior space sized to accommodate the reservoir;

a gasket positioned between the base rim and the lid rim when the base and the lid are in a closed relationship, the gasket forming a hermetic seal when compressed between the base rim and the lid rim;

one or more sealing elements each in closeable engagement with a corresponding one of the vent apertures such that the sealing element and corresponding vent aperture are operable between:

an open configuration in which the position of the sealing element relative to the corresponding vent allows fluid communication between the interior space and air outside the cartridge when the base and lid are in a closed relationship; and

a closed configuration in which the position of the sealing element relative to the corresponding vent does not allow fluid communication between the interior space and air outside the cartridge when the base and the cover are in a closed relationship, such that the interior space is sealed from air outside the cartridge when the base and the cover are in a closed relationship.

2. The lyophilization kit of item 1, further comprising one or more fasteners for securing the lid to the base, the fasteners configured to compress a gasket between the base edge and the lid edge to form an air-tight seal when the base and the lid are in a closed relationship.

3. The lyophilization kit of item 2, wherein the fastener comprises one or more latches.

4. The lyophilization kit of item 2, wherein at least one of the fasteners comprises a base pin, a cam lever, and a connecting arm.

5. The lyophilization kit of any one of clauses 1-4, wherein the base comprises aluminum.

6. The lyophilization kit of any one of clauses 1-5, wherein the top surface of the floor comprises one or more reservoir receiving portions formed therein, the reservoir receiving portions each having features that follow at least a bottom end of a reservoir to be received thereon and being configured to conduct heat between a base and a reservoir supported on the base.

7. The lyophilization kit of item 6, wherein:

the reservoirs each comprise a plurality of freeze-dried wells; and

the reservoir receiving portions each include a plurality of well receiving features that follow features of at least a bottom end of a freeze-dried well of a respective reservoir.

8. The lyophilization kit of any one of items 1-7, wherein:

each of said sealing elements being a flexible insert comprising a sealing cap and a body portion depending from said sealing cap, said body portion being in closable engagement with a corresponding one of said vent holes and having one or more vent slots formed therein; and

the sealing elements are each configured to be disposed in a corresponding one of the vent holes such that (i) when the sealing element and corresponding vent hole are in an open configuration, the interior space is in fluid communication with air outside the kit via the vent slot; and (ii) the interior space is not in fluid communication with air outside the cartridge when the sealing element and corresponding vent are in a closed configuration with the bottom surface of the sealing cap in sealing contact with the top surface of the top plate.

9. The lyophilization kit of item 8, wherein the closable engagement is a sliding engagement.

10. The lyophilization kit of item 1, wherein the closable engagement is a sliding engagement, a hinged engagement, a swinging engagement, or a rotational engagement.

11. The lyophilization kit of any one of items 1-10, further comprising one or more reservoir frames located on the top surface of the floor, the reservoir frames each being configured to support a plurality of reservoirs.

12. The lyophilization kit of item 11, wherein the reservoir frames each comprise aluminum.

13. A system for lyophilizing a substance, the system comprising:

a sealable enclosure defining a chamber;

the lyophilization nest of any of items 1-12 contained within the chamber, wherein the base and the lid are in a closed relationship, and wherein the interior space of the lyophilization nest comprises at least one reservoir containing a substance to be lyophilized; and

an upper shelf and a lower shelf contained within the chamber, wherein the lyophilization nest is positioned on the lower shelf, and wherein at least one of the upper shelf and the lower shelf is capable of automated movement that engages the upper shelf with the sealing element to change the sealing element and corresponding vent from the open configuration to the closed configuration.

14. The lyophilization system of item 13, wherein the upper shelf is configured to automatically move downward toward a top surface of a lid of the lyophilization nest.

15. The lyophilization system of clauses 13 or 14, wherein the reservoirs are each formed from a plastic material having a low moisture vapor transmission rate, and wherein the reservoirs each comprise a plurality of pores configured to be sealed.

16. A method for lyophilizing a substance, the method comprising the steps of:

(a) disposing a lyophilization nest on a lower shelf contained within a lyophilizer chamber, the lyophilization nest supporting one or more reservoirs contained within an interior space of the lyophilization nest, at least one of the reservoirs containing a substance to be lyophilized, wherein the interior space of the lyophilization nest is in fluid communication with air outside the nest through one or more vents extending through a top plate of a lid of the lyophilization nest, and wherein the vents each have a sealing element in closeable engagement therewith;

(b) closing a chamber containing the lyophilization nest;

(c) creating lyophilization conditions within the chamber for a time sufficient to lyophilize a substance contained in at least one of the reservoirs;

(d) when the chamber is closed, moving at least one of a lower shelf and an upper shelf contained within the chamber such that the upper shelf engages with the sealing element, thereby closing the vent hole, sealing the interior space of the lyophilization nest from air outside the nest, and stabilizing absolute humidity within the interior space of the lyophilization nest; and

(e) opening the chamber and removing the kit from the lyophilizer.

17. The method of clause 16, wherein the step of creating lyophilization conditions comprises creating a vacuum and a temperature below freezing within the chamber.

18. The method of clause 17, wherein the step of creating lyophilization conditions comprises creating a vacuum within the chamber and cycling between a first temperature below freezing and a second temperature below freezing, each of the first temperature and the second temperature being below the freezing temperature of the substance to be lyophilized.

19. The method of

clauses

16, 17, or 18, wherein after step (d), the absolute humidity within the interior space of the lyophilization nest is about 0.0 grams of water per cubic meter of air.

20. The method of

clauses

16, 17, or 18, wherein after step (d), the absolute humidity within the interior space of the lyophilization nest is less than 2.3 grams of water per cubic meter of air.

21. The method of item 20, wherein after step (d), the absolute humidity within the interior space of the lyophilization nest is less than 1.15 grams of water per cubic meter of air, more preferably less than 0.23 grams of water per cubic meter of air.

22. The method of any one of items 16-21, wherein the lyophilization nest is the lyophilization nest of any one of items 1-12, and wherein the base and the lid are in a closed relationship.

23. The method of any one of items 16-22, further comprising the steps of:

(f) after step (e), removing the lyophilization nest from the chamber;

(g) placing and removing the reservoir from the lyophilization nest; and

(h) the reservoirs are sealed to form an airtight seal between each of the plurality of apertures of each reservoir and the air outside the apertures of the sealed reservoirs.

24. The method of item 23, wherein steps (g) and (h) are performed in a space where humidity is not controlled.

25. The method of clause 23 or 24, wherein step (h) comprises securing a low moisture vapor permeable membrane to a top surface of each reservoir, thereby forming an airtight seal between the aperture and the air outside the aperture of the sealed reservoir.

26. The method of item 25, wherein the low moisture vapor permeable film is a laminate comprising an aluminum foil layer.

27. The method of item 25 or 26, wherein the low moisture vapor transmission film further comprises an adhesive.

28. The method of clause 25 or 26, wherein the low moisture permeable membrane further comprises a plastic liner for attaching the membrane to the top surface of each reservoir.

29. The method of any one of items 16-28, wherein, after step (e), the absolute humidity within the interior space of the lyophilization nest is maintained for at least 4 hours at less than 2.3 grams of water per cubic meter of air.

30. The method of item 16, wherein the closeable engagement is a sliding engagement of the sealing element with the vent.

31. The method of item 16, wherein the closable engagement is a hinged, swinging, or rotating engagement of the sealing element with the vent.

Claims

1. A reservoir for receiving one or more substances to be lyophilized, the reservoir comprising:

one or more wells, each well configured to hold a substance to be lyophilized and comprising an at least substantially hemispherical bottom interior surface; and

a sealable upper surface configured such that a seal can be applied thereto and the upper surface sealed such that the one or more apertures are each individually sealable and isolated from an uncontrolled environment external to the sealed reservoir,

wherein an external shape of at least a bottom of the reservoir is configured to follow and engage with one or more reservoir receiving portions of a lyophilization nest so as to maximize a surface contact area between the reservoir and the corresponding one or more reservoir receiving portions, an

Wherein the reservoir comprises a low vapor permeability material configured to inhibit migration of water vapor through the material.

2. The reservoir of claim 1, wherein said reservoir is a single-well reservoir comprising one well, or a multi-well reservoir comprising a plurality of wells.

3. The reservoir of claim 1, wherein said low vapor permeability material comprises a cyclic olefin copolymer or polypropylene.

4. The reservoir of claim 1, further comprising a seal attached to the sealable upper surface, wherein the seal comprises an adhesive low moisture vapor transmission material secured to the sealable upper surface of the reservoir.

5. The reservoir of claim 4, wherein the adhesive low moisture vapor transmission material is a film, and wherein the film is a laminate structure comprising a layer of aluminum foil.

6. The reservoir of claim 1, wherein the one or more wells each have an associated sealing cap configured to be secured within the well to provide an airtight seal around a perimeter of the well.

7. The reservoir of claim 1, wherein the one or more wells each comprise a flat bottom exterior surface, a V-shaped bottom exterior surface, or an at least substantially hemispherical bottom exterior surface corresponding to the at least substantially hemispherical bottom interior surface.

8. The reservoir of claim 1, wherein the one or more substances to be lyophilized comprise at least one of: pure chemicals; a chemical mixture; biological samples such as cells, cell extracts or tissues; biological agents such as nucleic acids, enzymes, antibodies, and proteins; labels such as fluorophores; and a reaction mixture.

9. The reservoir of claim 2, comprising a 6, 12, 24, 96, 384, or 1536 well multi-well reservoir.

10. The reservoir of claim 1, wherein the volume of each of said one or more wells ranges from about 300 μ L to about 400 μ L.

11. The reservoir of claim 1, wherein the volume of each of said one or more wells ranges from about 1nL to 10 mL.

12. A lyophilization kit for preparing a lyophilized material, the kit comprising:

a base comprising a bottom plate having a base edge extending upwardly from a perimeter thereof, the bottom plate having a top surface adapted to support at least one reservoir of claim 1;

an interior space defined by the base and the lid when the base and the lid are in a closed relationship, the interior space sized to accommodate the at least one reservoir; and

one or more sealing elements each in closeable engagement with a corresponding one of the vents such that the sealing element and corresponding vent are operable between:

13. The lyophilization kit of claim 12, further comprising one or more reservoir frames located on a top surface of the floor, the reservoir frames each configured to support the at least one reservoir, wherein the reservoir frames each comprise a metallic material, the metallic material being aluminum.

14. A method for sealing a lyophilized substance in a reservoir, the method comprising:

preparing a lyophilized material in a reservoir comprising one or more pores, wherein preparing the lyophilized material in the reservoir comprises:

placing a substance to be lyophilized in one or more wells of the reservoir;

positioning the reservoir within a lyophilization nest that includes an interior space in fluid communication with air outside the nest through one or more vent holes extending through a top panel of a lid of the lyophilization nest, and wherein the vent holes each have a sealing element in closeable engagement therewith; and is

Placing the lyophilization nest within a chamber of a lyophilizer and creating lyophilization conditions within the chamber for a time sufficient to lyophilize a substance contained in a reservoir;

removing the lyophilization nest from the chamber of the lyophilizer;

placing and removing the reservoir from the lyophilization nest; and is

Securing a low moisture vapor transmission membrane to a top surface of the reservoir such that the one or more apertures are each individually sealable and isolated from an uncontrolled environment external to the sealed reservoir,

wherein the low moisture vapor transmission film is a laminate structure comprising an aluminum foil layer.

15. The method of claim 14, wherein securing the low moisture vapor transmission membrane to the top surface of the reservoir is performed in a space without controlling humidity.

16. The method of claim 14, comprising placing about 75-200 μ L of the material to be lyophilized in the one or more wells of the reservoir.

17. The method of claim 14, wherein the low moisture vapor transmission film further comprises an adhesive; or wherein the low moisture vapor transmission film further comprises a plastic liner for affixing the film to the top surface of the reservoir.

18. A system for maximizing heat transfer within a lyophilization nest, the system comprising:

a lyophilization kit, the kit comprising:

a lid comprising a top panel having a lid edge extending downwardly from a perimeter thereof, the top panel having one or more vent holes extending therethrough; and

a base comprising a floor having a base edge extending upwardly from a perimeter thereof and defining a base interior comprising one or more reservoir-receiving portions configured to support one or more reservoirs therein,

wherein each reservoir comprises:

a sealable upper surface configured such that a seal can be applied thereto and the upper surface is sealed such that the one or more apertures are each individually sealable and isolated from an uncontrolled environment external to the sealed reservoir, an

Wherein an exterior shape of at least a bottom portion of each reservoir is configured to follow and engage with one or more reservoir-receiving portions in the interior of the base to maximize a surface contact area between the reservoir and the corresponding one or more reservoir-receiving portions, thereby maximizing heat transfer between the reservoir and the corresponding one or more reservoir-receiving portions.

19. The system of claim 18, further comprising a seal attached to the sealable upper surface, wherein the seal comprises an adhesive low moisture vapor permeable material secured to the sealable upper surface of the reservoir.

20. The system of claim 18, wherein the reservoir comprises a material configured to be low vapor permeability that inhibits migration of water vapor through the material.

21. The system of claim 18, wherein the one or more reservoir-receiving portions comprise a material having a high heat transfer coefficient and may be in sufficient contact with the base such that heat transfer between the one or more reservoir-receiving portions and the base is maximized.

22. The system of claim 21, wherein the one or more reservoir-receiving portions are formed from a material that is a unitary material with the base such that the one or more reservoir-receiving portions are integral with the base.

23. The system of claim 21, wherein the one or more reservoir-receiving portions are removably secured to the base.

24. The system of claim 21, wherein the one or more reservoir-receiving portions comprise a metallic material, the metallic material being aluminum.